A solid-state culture system using a cellulose carrier containing defined medium as a useful tool for investigating characteristics of koji culture

The purpose of this article is to establish a new solid-state culture system that uses a defined medium and allows control of the chemical factors in the medium in solid-state cultivation. To achieve this purpose, we produced a solid substrate, in which the defined medium was packed in a cellulose carrier with agar, and named it "cellulose agar cube (CA-cube)". In order to obtain growth conditions similar to koji culture, we then determined the moisture content of the CA-cube. Five runs of column reactor operations were carried out at various moisture contents of the CA-cube in the range of 50-90%. A very similar mycelial morphology to that of koji culture was observed on the surface of CA-cube with moisture content lower than 63%. The glucoamylase production and the production ratio of glucoamylase to alpha-amylase (G/A ratio) were both increased exponentially with decreasing moisture content and reached 237.5 units per g dry solid and 13.59, respectively, at the moisture content of 50%, which are almost comparable to those of koji culture. In addition, the glucoamylase "glaB", which is mainly produced in koji culture, was observed in the CA-cube culture with low moisture content. These results indicate that growth conditions similar to koji culture could be attained by the column reactor culture with the CA-cube.     

Sugar beet pectin as a natural carrier for curcumin,a water-insoluble bioactive for food and beverage enrichment: Formation and characterization

Food and beverages enrichment with water-insoluble health-promoting nutraceuticals is important, but technologically challenging. Sugar beet pectin (SBP) is a natural dietary fiber with high emulsifying capacity. However, its potential as a natural encapsulator of hydrophobic nutraceuticals for beverage enrichment, has hardly been explored. Curcumin is a potent antioxidant with numerous attributed health benefits, but very low aqueous-solubility. We herein explored SBP as a carrier for solubilization and protection of curcumin (CUR). Using spectrofluorimetry, the CUR-SBP binding constant determined was (6.74 ± 0.5) ∙ 10⁵M⁻¹. As CUR:SBP molar ratio increased from 14:1 to 140:1, CUR encapsulation capacity increased from 14.5 ± 0.8 to 127.4 ± 0.4 mg _{CUR/g SBP}, and encapsulation efficiency moderately decreased from ~100% to 86 ± 7%. The encapsulation in SBP dramatically decreased CUR particle size, from >17 μm to <0.5 μm, in average, and conferred substantial protection to CUR during simulated shelf-life, decreasing the decay rate constant ~7 fold. Therefore, SBP is a potent natural encapsulator for hydrophobic nutraceuticals for food and even clear beverage enrichment.     

Microbiological results from milk samples obtained premilking and postmilking for the diagnosis of bovine intramammary infections.

Bacteriological culture results were compared between 336 pairs of quarter milk samples collected premilking and postmilking. Using a positive result on either premilking or postmilking samples as the definitive diagnosis, premilking sampling sensitivity was 91% for Staphylococcus aureus, 91% for coagulase-negative staphylococci, and 97% for Streptococcus other than agalactiae. Postmilking sampling sensitivities were 81, 45, and 58%, respectively, for the same pathogens. Requiring both premilking and postmilking samples for the definitive diagnosis, specificities were 92, 86, and 95% for premilking sampling alone and 96, 98, and 99% for postmilking sampling alone. Such differences in specificity would result in major differences in predictive value of a positive culture for herds with a low prevalence. Multiple isolates were significantly more common from premilking samples.     

Stability of betacyanin from red pitahaya (Hylocereus polyrhizus) and its potential application as a natural colourant in milk

The effects of temperature (30, 50, 85 and 100 °C) treatment and 10 weeks of refrigerated storage (4 °C) on the betacyanin content and colour stability of betacyanins from red pitahaya (Hylocereus polyrhizus) were investigated and compared to red beet (Beta vulgaris; E‐162) which was used as a control. Temperature treatment at 50 and 85 °C caused a greater reduction in betacyanin content (BC) in E‐162 compared to red pitahaya by inducing changes in betacyanin profile. The 10 weeks of refrigerated storage at 4 °C induced colour changes in betacyanins from red pitahaya and E‐162. In milk, betacyanins from red pitahaya presented a lower loss of BC and total colour changes (ΔE*) compared to E‐162. The microbial onset in milk containing betacyanins from red pitahaya and E‐162 was delayed up to day 5 of refrigerated storage at 4 °C compared to day 3 in plain milk. Milk containing betacyanins from red pitahaya showed better colour acceptability (3.89 ± 1.89) compared to E‐162 (6.10 ± 1.71). Hence, betacyanins from red pitahaya may be a potential natural functional colourant to simulate strawberry colour in milk.     

Proteolysis in milk: the significance of proteinases originating from milk leucocytes and a comparison of the action of leucocyte, bacterial and natural milk proteinases on casein

The caseinolytic activities at pH 6.8 of polymorphonuclear (PMN) and mononuclear leucocyte homogenates (equivalent to a level of 10(6) cells/ml milk) were less than the levels of natural milk proteinase activity found in milk from healthy cows. Bulk milks contained approximately 4 times more milk proteinase activity than the composite milks from individual healthy cows. Isolated blood leucocytes, when added to raw milk of good bacteriological quality and stored at 5 degrees C, did not readily degenerate and had no detectable effect on the milk proteins even when these cells were completely disrupted by homogenization of the milk. Pasteurization of milk which contained leucocytes caused loss of cell vitality. Extracellular proteinases of psychrotrophic bacteria growing in milk were not detected until the early stationary phase of growth. The total viable count at which this occurred varied greatly. Proteinase production by a pure culture of Pseudomonas fluorescens was not detected in milk stored at 5 degrees C until a viable count of approximately 10(9) colony forming units (c.f.u.)/ml was obtained, whilst normal bulk milks stored at 5 degrees C produced detectable levels of extracellular proteinase(s) when the psychrotrophic flora reached 10(7)-10(8) c.f.u./ml. Casein proteolysis by PMN and mononuclear leucocyte homogenates resulted in similar polypeptide maps, but plasmin and bacterial proteinase isolated from a strain of Serratia marcescens resulted in polypeptide maps different from each other and from that produced by the leucocyte proteinase(s). The rate of proteolysis of caseins by the different proteinase sources appeared to be in the order alpha s1- greater than beta- greater than greater than kappa-casein for the leucocyte extracts, beta- greater than alpha s1- greater than greater than greater than kappa-casein for bovine plasmin and beta- approximately kappa- greater than alpha s1-casein for for S. marcescens proteinase.     

Pressurisation of raw skim milk and of a dispersion of phosphocaseinate at 9 degrees C or 20 degrees C: effects on the distribution of minerals and proteins between colloidal and soluble phases

The effects of high pressure treatments (100-300 MPa; 15 min; 9 degrees C or 20 degrees C) on the distribution of minerals and proteins of raw skim milk (RSM) and of a dispersion of industrial phosphocaseinate (PC) were studied after separation of the micellar and soluble phases by ultracentrifugation (UCF). Whatever the temperature of high pressure treatments, the pressure-induced dissociation of the casein micelles was accompanied by calcium (Ca), phosphorus (P) and casein release from the micelles. The released Ca and P were or became bound to soluble proteins since progressive increases in Ca and P concentrations were observed in the UCF supernatants of RSM and of the PC dispersion but not in the ultrafiltrates from these UCF supernatants (free of soluble proteins). Simultaneously, alpha(S1-), alpha(S2-), beta- and kappa-caseins were progressively released from the micelles, as seen by electrophoretic analysis. The pressure-induced solubilisation of alpha(S1-) and alpha(S2-)caseins, essentially located in the core of the micelles, suggests that high pressure de-stabilized micelles including their internal structure.     

Volatile organic compounds from the interaction between Fusarium verticillioides and maize kernels as a natural repellents of Sitophilus zeamais

Maize kernels are exposed to Sitophilus zeamais attack and Fusarium verticillioides infestation during storage, which can result in product deterioration and economic losses. The objective of this study was to evaluate the involvement of the Volatile Organic Compounds (VOCs) emitted by the fungi-corn system in grain-insect interactions. Volatiles emitted by healthy maize kernels were different from those emitted by fungal infected kernels, with the latter being enriched by alcohols, ketones and sesquiterpenes, which were considered early indicators of fungal contamination. The results demonstrated that the kernels exposed to the fungal VOCs and their pure compounds (1-octen-3-ol and 3-octanol) were less attractive and less damaged by S. zeamais than controls. In addition to compound adsorption, other processes may have caused the protective effect of exposed kernels against insect damage. This is the first contribution of the role of the fungal volatiles on the behavior of S. zeamais, and could provide an important contribution to the conservation of stored grains and pest management and an early indicator of fungal contamination.     

Antimicrobial peptides generated from milk proteins: a survey and prospects for application in the food industry. A review

Milk proteins constitute a natural reservoir of bioactive peptides with physiological and/or antimicrobial properties, the release of which requires hydrolysis of the precursor molecules by digestive proteases or by fermentation with proteolytic micro‐organisms. Depending on the digestive or microbial proteases used, an array of bioactive peptides would be released either from caseins or whey proteins, but only a small part of these peptides has so far been identified and characterised with respect to their antimicrobial activity. The antimicrobial peptides known thus far have proven to be potent inhibitors to the growth of a wide range of undesirable micro‐organisms of health or spoilage significance. Nevertheless, previous research work has largely been oriented towards their possible application in medicine, which has hindered their high potential as food‐grade biopreservatives and/or as supplements in functional foods. This review attempts to study the literature pertaining to antimicrobial peptides derived from major milk proteins (caseins, α‐lactalbumin and β‐lactoglobulin) upon hydrolysis either by digestive proteases or by fermentation with proteolytic lactic acid bacteria. Their possible application in the food industry and their mechanism of action will also be discussed. Reference antimicrobial peptides produced by living micro‐organisms as innate immune defence components against microbial infections will occasionally be invoked for comparison purposes.     

Fluorescence and color as markers for the Maillard reaction in milk–cereal based infant foods during storage

Free and total fluorescence compounds and color formation were measured in three different milk–cereal based infant foods stored at 25, 30 and 37 °C for 9 months to evaluate the advanced and final stages of the Maillard reaction. Milk–cereal infant foods containing honey (B) or fruits (C) had fluorescent values higher than sample (A) without them. This difference could be ascribed to the higher monosaccharide (fructose and/or glucose) content of (B) and (C), which could increase susceptibility to the Maillard reaction. However, for color increase (ΔE), no significant differences (p < 0.05) among the three types of samples were found. During the storage period, a gradual increase in fluorescence and color was observed, and statistically significant differences among the three temperatures studied were detected, the values being greater at 37 °C than at 30 °C and 25 °C.     

In vitro and in vivo evidence for a satiating effect of fish protein hydrolysate obtained from blue whiting (Micromesistius poutassou) muscle

To find appetite suppressive molecules derived from fish protein hydrolysates, both in vitro and in vivo experiments were performed in order to demonstrate that hydrolysates produced from blue whiting muscle (BWMH) possess satiating properties. Here we demonstrated for the first time that a protein hydrolysate obtained from marine source was able to enhance cholecystokinin (CCK) and glucagon-like peptide-1 (GLP-1) secretion in STC-1 cell line. To demonstrate that these in vitro activities also exist in vivo, we investigated the effect of BWMH preload administration in rats and its repercussion on food intake and metabolic plasma marker levels. Results showed that BWMH reduced the short term food intake which was correlated to an increase in the CCK and GLP-1 plasma levels. Moreover it was demonstrated that the chronic administration of BWMH led to a decrease in the body weight gain.     

Proteolysis of milk proteins by AprX,an extracellular protease identified in Pseudomonas LBSA1 isolated from bulk raw milk,and implications for the stability of UHT milk

UHT milk made from milk contaminated by Pseudomonas LBSA1 destabilised during storage. Sedimentation of UHT milk was observed; zeta potential of casein micelles decreased, while contents of non-casein nitrogen and non-protein nitrogen increased. Pseudomonas LBSA1 produced an extracellular protease that hydrolysed caseins but not whey proteins; this was identified as AprX, a thermoresistant protease belonging to the serralysin family. This protease showed a broad range of pH activity (pH 6 to pH 10) and an optimal temperature of activity of 40 °C. Peptides released from purified α_S1-, β- and κ-caseins were determined by tandem mass spectrometry. The identified cleavage sites did not reveal a strong specificity of the extracellular protease. However, the presence of basic or aromatic amino acid residues in the P1 position had a positive influence on cleavage in comparison with acidic amino acid residues or proline.     

CHROMagar Candida medium as a practical tool for the differentiation and presumptive identification of yeast species isolated from salads

CHROMagar Candida medium was used to study the diversity of yeast biota of salad samples, and to presumptively identify the isolates. This medium was originally developed for the selective isolation and presumptive identification of some clinically important yeast species such as Candida albicans, Candida tropicalis, Candida krusei, and Candida glabrata on the basis of differences in colour and surface of colonies. Ninety three yeast strains representing 33 species from the culture collection and 39 fresh isolates from different mayonnaise-based mixed salads showed a wide range of hue of colony colours ranging from white to yellow, orange, red, pink, purple, blue, green, etc., as well as different morphological appearances on the CHROMagar Candida medium. Therefore, CHROMagar Candida medium facilitates the detection of mixtures of yeast species from different samples on a single isolation plate and this medium can be a practical method for the differentiation and rapid presumptive identification of many yeast species occurring frequently in different kind of foods.     

Assessing the effectiveness of colloidal microcrystalline cellulose as a suspending agent for black and white liquid dyes

This study aimed to determine the effect of the concentration (1, 5, or 8 wt%) of colloidal microcrystalline cellulose (MCCS) as suspending agent on the long-term stability and rheology of the vegetable carbon (C) and calcium carbonate (CC) suspended in water–glycerine mixture. These suspensions as ready-to-use black and white liquid dyes containing 10 wt% C or CC were assessed for particle size, Turbiscan stability index (TSI), steady shear, thixotropy and dynamic viscoelasticity. After 370 days, the black dyes showed excellent stability (TSI ≪ 0.5) at 4 °C and 25 °C already at 1 wt% MCCS, while the white dyes showed reasonable stability (TSI 1–3) only at 5 and 8 wt% MCCS. Dye formulations exhibited a relatively liquid-like viscoelastic behaviour, while they showed extremely shear-thinning behaviour with a network structure dependent on the concentration of the MCCS as exhibited by the increase in thixotropy and the existence of the measurable yield stress.     

Identification of kappa-casein genotype in Holstein sires: a comparison between analysis of milk samples from daughters and direct analysis of semen samples from sires by polymerase chain reaction

Two different methods were used to determine kappa-casein genotypes of Holstein sires. In the earlier procedure, genotypes of sires were deduced by analyzing frequency distribution data of kappa-casein variants obtained through typing of milk samples from daughters by electrophoresis. The second method involved direct analysis of DNA obtained from semen samples of the sires. The polymerase chain reaction was used to amplify a 99-bp region from the kappa-casein gene that contains nucleotide substitutions that are diagnostic of variants A and B. Identity of the amplified product was confirmed by sequencing. Results obtained by both methods of genotyping were similar. For the 42 sires that were analyzed by both methods, the distribution of kappa-casein genotypes were 31 AA, 11 AB, and 0 BB. The frequency of B allele for kappa-casein in the sire population studied was lower than in larger Holstein cow populations. Due to the high demand for kappa-casein B milk by the dairy industry, it might be advantageous to increase this allele in the dairy cattle population by identifying sires with homozygous kappa-casein B and using them more frequently as service sires.